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Natural Fe isotope fractionation in an intestinal Caco-2 cell line model†
Journal of Analytical Atomic Spectrometry ( IF 3.1 ) Pub Date : 2017-04-28 00:00:00 , DOI: 10.1039/c7ja00090a María R. Flórez 1, 2, 3, 4, 5 , Yulia Anoshkina 1, 2, 3, 4, 5 , Marta Costas-Rodríguez 1, 2, 3, 4, 5 , Charlotte Grootaert 2, 4, 5, 6, 7 , John Van Camp 2, 4, 5, 6, 7 , Joris Delanghe 4, 5, 8, 9 , Frank Vanhaecke 1, 2, 3, 4, 5
Journal of Analytical Atomic Spectrometry ( IF 3.1 ) Pub Date : 2017-04-28 00:00:00 , DOI: 10.1039/c7ja00090a María R. Flórez 1, 2, 3, 4, 5 , Yulia Anoshkina 1, 2, 3, 4, 5 , Marta Costas-Rodríguez 1, 2, 3, 4, 5 , Charlotte Grootaert 2, 4, 5, 6, 7 , John Van Camp 2, 4, 5, 6, 7 , Joris Delanghe 4, 5, 8, 9 , Frank Vanhaecke 1, 2, 3, 4, 5
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In this work, Fe isotopic analysis of samples obtained from an in vitro intestinal model was performed via multi-collector ICP-mass spectrometry (MC-ICP-MS) to evaluate the isotope fractionation accompanying Fe uptake and transport mechanisms at a cellular level. The Caco-2 cell line has been used, after cell differentiation, as an enterocyte model and a bi-cameral experimental setup has been developed and optimized for stimulating intracellular Fe fluxes. An Fe : ascorbic acid mixture with a molar ratio of 1 : 5 was used as a source of non-heme bioavailable Fe. Good experimental repeatability and reproducibility were attained with low blank contribution levels, allowing precise and reliable Fe isotope ratio results. Both Fe absorption and transport processes were accompanied by Fe isotope fractionation in favor of the lighter isotopes. After 3 hours of exposure, the isotopic composition of the apical solution and the cells did not significantly differ from that of the original solution added to the cells. After 24 hours of exposure, the trend observed was towards a light Fe isotopic composition in the cells, whereas the apical solutions were enriched in the heavier isotopes. These results were in good agreement with previous in vivo and ex vivo findings. An overall increase in delta Fe values of the cell layers exposed to Fe treatment relative to the corresponding values for the untreated cells also seems to support the assumption of a preferential accumulation of heavy isotopes in enterocyte ferritin. The consistency of the results obtained supports the usefulness of in vitro cell culture models as an interesting complementary tool for studying Fe metabolic pathways at the intestinal level.
中文翻译：
肠道Caco-2细胞系模型中的天然铁同位素分馏†
在这项工作中，对通过体外肠道模型获得的样品进行的铁同位素分析是通过多收集器ICP-质谱（MC-ICP-MS）在细胞水平上评估伴随Fe吸收和传输机制的同位素分级。在细胞分化后，已经将Caco-2细胞系用作肠细胞模型，并且已经开发并优化了双子实验装置来刺激细胞内的铁通量。摩尔比为1：5的Fe：抗坏血酸混合物被用作非血红素可生物利用的Fe的来源。在空白贡献水平较低的情况下，可以获得良好的实验可重复性和可重复性，从而获得精确可靠的铁同位素比结果。Fe的吸收和输运过程都伴随着Fe同位素分级分离，有利于更轻的同位素。暴露3小时后，顶端溶液和细胞的同位素组成与添加到细胞中的原始溶液的同位素组成没有显着差异。暴露24小时后，观察到的趋势是细胞中的Fe同位素组成变轻，而顶端溶液中富集了较重的同位素。这些结果与以前的结果非常吻合体内和离体发现。相对于未处理的细胞的相应值，暴露于铁处理的细胞层的δ铁值的总体增加似乎也支持了重同位素在肠细胞铁蛋白中优先积累的假设。获得的结果的一致性支持体外细胞培养模型作为研究肠道水平Fe代谢途径的有趣补充工具的有用性。
更新日期：2017-04-28
中文翻译：
肠道Caco-2细胞系模型中的天然铁同位素分馏†
在这项工作中，对通过体外肠道模型获得的样品进行的铁同位素分析是通过多收集器ICP-质谱（MC-ICP-MS）在细胞水平上评估伴随Fe吸收和传输机制的同位素分级。在细胞分化后，已经将Caco-2细胞系用作肠细胞模型，并且已经开发并优化了双子实验装置来刺激细胞内的铁通量。摩尔比为1：5的Fe：抗坏血酸混合物被用作非血红素可生物利用的Fe的来源。在空白贡献水平较低的情况下，可以获得良好的实验可重复性和可重复性，从而获得精确可靠的铁同位素比结果。Fe的吸收和输运过程都伴随着Fe同位素分级分离，有利于更轻的同位素。暴露3小时后，顶端溶液和细胞的同位素组成与添加到细胞中的原始溶液的同位素组成没有显着差异。暴露24小时后，观察到的趋势是细胞中的Fe同位素组成变轻，而顶端溶液中富集了较重的同位素。这些结果与以前的结果非常吻合体内和离体发现。相对于未处理的细胞的相应值，暴露于铁处理的细胞层的δ铁值的总体增加似乎也支持了重同位素在肠细胞铁蛋白中优先积累的假设。获得的结果的一致性支持体外细胞培养模型作为研究肠道水平Fe代谢途径的有趣补充工具的有用性。
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